Bovine cornea, sclera, iris, ciliary body, choroid, zonular fibers, lens capsule, lens nucleus, vitreous body, and retina were investigated for collagen content and type. Cornea, sclera, iris, ciliary body, choroid, lens capsule, and vitreous body contain hydroxyproline, whereas in zonular fibers, lens nucleus, and retina no hydroxyproline was detectable. Preparative isolation of collagen was achieved by digestion of the different eye tissues with pepsin. The pepsin-solubilized collagen was separated by differential salt precipitation into different collagen types. The polyacrylamide gel electrophoresis of the pepsin-solubilized collagens revealed type I collagen in cornea, sclera, iris, ciliary body, and choroid. As well as type I collagen, type III collagen was isolated from cornea, sclera, and uveal tissues. The identification of types I and III collagen was supported by the CNBr-derived peptides of these collagens. Lens capsule collagen consisted mainly of type IV collagen. Zonular fibers contained no hydroxyproline but when examined by polyacrylamide gel electrophoresis, a band migrating in the alpha-position of collagen was observed. Polyacrylamide gel electrophoresis of both the pepsin-solubilized component and the CNBr-derived peptides of vitreous body protein showed no relation to any of the four common collagen types.